Submitted to: Proceedings of Pittcon Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 3/15/2012
Publication Date: 3/15/2012
Citation: Roberts, E., Boudreau, B., Larson, T.M., Brown, D.W., Mcquade, K., Remsen, E.E. 2012. Quantitative and Qualitative Analysis of Biomarkers in Fusarium verticillioides. Proceedings of Pittcon Meeting. Interpretive Summary:
Technical Abstract: In this study, a combination HPLC-DART-TOF-MS system was utilized to identify and quantitatively analyze carbohydrates in wild type and mutant strains of Fusarium verticillioides. Carbohydrate fractions were isolated from F. verticillioides cellular extracts by HPLC using a cation-exchange size-exclusion column and permethylated for DART-TOF-MS analysis. Quantitative analysis was performed with the HPLC system's differential refractive index detector after calibration with standards of known concentration. This approach allows for multiple carbohydrates to be identified and quantified in a single cellular extract. This method is being used to characterize the carbohydrate profile in F. verticillioides and examine how deleting genes involved in trehalose metabolism affects this profile. Significant amounts of all five carbohydrates tested were detected in the wild type strain, with mannitol being the most abundant. Deletion of the trehalose-6-phosphate synthase gene causes F. verticillioides to be compromised in its ability to make trehalose. Deletion of either the trehalose phosphate phosphatase or trehalose phosphorylase gene results in a dramatic increase in mannitol and a concomitant decrease in arabitol.